LOCUS       sequence-274635-        3733 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Drug Resistance pDONR221SAT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3733)
  AUTHORS   Duncan SM, Myburgh E, Alves-Ferreira EV, Mottram JC
  TITLE     DiCre-Based Inducible Disruption of Leishmania Genes.
  JOURNAL   Methods Mol Biol. 2019;1971:211-224. doi: 
            10.1007/978-1-4939-9210-2_10.
  PUBMED    30980305
REFERENCE   2  (bases 1 to 3733)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Methods Mol
            Biol."; date: "2019"; volume: "1971:211-224. doi"; pages: " 
            10.1007/978-1-4939-9210-2_10"
COMMENT     Sequence Label: pGL2206
FEATURES             Location/Qualifiers
     source          1..3733
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     98..115
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(274..301)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(393..479)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     528..550
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     542..559
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     543..559
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    575..674
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(1671..1682)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(1864..1963)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1980..1999)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1981..1999)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2004..2020)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2004..2020)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2122..2141)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2133..2942
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2200..2219)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3089..3677
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3578..3597
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
       61 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
      121 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
      181 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg
      241 taccgctagc caggaagagt ttgtagaaac gcaaaaaggc catccgtcag gatggccttc
      301 tgcttagttt gatgcctggc agtttatggc gggcgtcctg cccgccaccc tccgggccgt
      361 tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc
      421 gacaaacaac agataaaacg aaaggcccag tcttccgact gagcctttcg ttttatttga
      481 tgcctggcag ttccctactc tcgcgttaac gctagcatgg atgttttccc agtcacgacg
      541 ttgtaaaacg acggccagtc ttaagctcgg gccccaaata atgattttat tttgactgat
      601 agtgacctgt tcgttgcaac aaattgatga gcaatgcttt tttataatgc caactttgta
      661 caaaaaagca ggctgtacac acgtacacac acacacacac acacacacac acacacacac
      721 acacgtacac acgtacacac acgtgcgcat gcttgacgca tacgcgacga attcgaaagc
      781 tcacctcatt cctccctcct cacaccatca tcggcatcca tagagacacg tgcgcgtaga
      841 acgatacagc tcacgcgcac agagaggcgt gttctggtcg tgcgcatcat cggcacagca
      901 ctggcggacg aaactcgcac acaggcacgc cgcctccttt cacccgtcat agatagttga
      961 attagacgcc ctcctcctcc ctcatcatcg ccgtcgtcat ccgggtccga gcactagtga
     1021 tgaagatttc ggtgatccct gagcaggtgg cggaaacatt ggatgctgag aaccatttca
     1081 ttgttcgtga agtgttcgat gtgcacctat ccgaccaagg ctttgaacta tctaccagaa
     1141 gtgtgagccc ctaccggaag gattacatct cggatgatga ctctgatgaa gactctgctt
     1201 gctatggcgc attcatcgac caagagcttg tcgggaagat tgaactcaac tcaacatgga
     1261 acgatctagc ctctatcgaa cacattgttg tgtcgcacac gcaccgaggc aaaggagtcg
     1321 cgcacagtct catcgaattt gcgaaaaagt gggcactaag cagacagctc cttggcatac
     1381 gattagagac acaaacgaac aatgtacctg cctgcaattt gtacgcaaaa tgtggcttta
     1441 ctctcggcgg cattgacctg ttcacgtata aaactagacc tcaagtctcg aacgaaacag
     1501 cgatgtactg gtactggttc tcgggagcac aggatgacgc ctaaggatcc tctagtagtg
     1561 atcccgcccg cacccgcgcg cgctcaggcc gcgtgtcgtc gtgctctttc catttttttt
     1621 ttgtgtgtgg tggtgatttg agctgctctc cgttgtgtgc tggggaccct ccttccctcg
     1681 atctcctcgt gcggggtctc cgacgccgca gacgcggtgc gtgcgagcgt gcacgtggct
     1741 gtcgccgtgg ctgtagttgc gagcggagag agagagagag ggagggaggg agggcagagg
     1801 gcagaggaca tgcgggtggg aacgtgcacc ggcctcgtct cacgcagctg gagcccacga
     1861 atgacccagc tttcttgtac aaagttggca ttataagaaa gcattgctta tcaatttgtt
     1921 gcaacgaaca ggtcactatc agtcaaaata aaatcattat ttgccatcca gctgatatcc
     1981 cctatagtga gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc
     2041 aaaatctctg atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt
     2101 ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtcga
     2161 ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata
     2221 atgtcgggca atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt
     2281 tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac
     2341 taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg
     2401 atgatgcatg gttactcacc actgcgatcc ccggaaaaac agcattccag gtattagaag
     2461 aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc
     2521 attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg
     2581 cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg
     2641 gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt
     2701 cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa
     2761 taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc
     2821 tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg
     2881 gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct
     2941 aatcagaatt ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggacg
     3001 gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag
     3061 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3121 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3181 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     3241 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3301 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3361 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     3421 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3481 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     3541 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     3601 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     3661 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     3721 ggccttttgc tca
//
